Advertisement

Science China Life Sciences

, Volume 62, Issue 12, pp 1670–1682 | Cite as

Supercritical fluid-assisted controllable fabrication of open and highly interconnected porous scaffolds for bone tissue engineering

  • Hanxiao Tang
  • Ranjith Kumar Kankala
  • Shibin Wang
  • Aizheng ChenEmail author
Research Paper
  • 45 Downloads

Abstract

Recently tremendous progress has been evidenced by the advancements in developing innovative three-dimensional (3D) scaffolds using various techniques for addressing the autogenous grafting of bone. In this work, we demonstrated the fabrication of porous polycaprolactone (PCL) scaffolds for osteogenic differentiation based on supercritical fluid-assisted hybrid processes of phase inversion and foaming. This eco-friendly process resulted in the highly porous biomimetic scaffolds with open and interconnected architectures. Initially, a 23 factorial experiment was designed for investigating the relative significance of various processing parameters and achieving better control over the porosity as well as the compressive mechanical properties of the scaffold. Then, single factor experiment was carried out to understand the effects of various processing parameters on the morphology of scaffolds. On the other hand, we encapsulated a growth factor, i.e., bone morphogenic protein-2 (BMP-2), as a model protein in these porous scaffolds for evaluating their osteogenic differentiation. In vitro investigations of growth factor loaded PCL scaffolds using bone marrow stromal cells (BMSCs) have shown that these growth factor-encumbered scaffolds were capable of differentiating the cells over the control experiments. Furthermore, the osteogenic differentiation was confirmed by measuring the cell proliferation, and alkaline phosphatase (ALP) activity, which were significantly higher demonstrating the active bone growth. Together, these results have suggested that the fabrication of growth factor-loaded porous scaffolds prepared by the eco-friendly hybrid processing efficiently promoted the osteogenic differentiation and may have a significant potential in bone tissue engineering.

Keywords

supercritical foaming polycaprolactone bone tissue engineering osteogenic differentiation bone morphogenic protein-2 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (U1605225, 31570974, and 31470927), the Public Science and Technology Research Funds Projects of Ocean (201505029), the Promotion Program for Young and Middle-aged Teacher in Science and Technology Research of Huaqiao University (ZQN-PY107) and the Program for Innovative Research Team in Science and Technology in Fujian Province University.

Compliance and ethics The author(s) declare that they have no conflict of interest.

Supplementary material

Supplementary material, approximately 15.1 MB.

References

  1. An, J., Teoh, J.E.M., Suntornnond, R., and Chua, C.K. (2015). Design and 3D printing of scaffolds and tissues. Engineering 1, 261–268.Google Scholar
  2. Autissier, A., Le Visage, C., Pouzet, C., Chaubet, F., and Letourneur, D. (2010). Fabrication of porous polysaccharide-based scaffolds using a combined freeze-drying/cross-linking process. Acta Biomater 6, 3640–3648.PubMedGoogle Scholar
  3. Brydone, A.S., Meek, D., and Maclaine, S. (2010). Bone grafting, orthopaedic biomaterials, and the clinical need for bone engineering. Proc Inst Mech Eng H 224, 1329–1343.PubMedGoogle Scholar
  4. Cabraja, M. and Kroppenstedt, S. (2012). Bone grafting and substitutes in spine surgery. J Neurosurg Sci 56, 87–95.PubMedGoogle Scholar
  5. Cai, Y., Tong, S., Zhang, R., Zhu, T., and Wang, X. (2018). In vitro evaluation of a bone morphogenetic protein-2 nanometer hydroxyapatite collagen scaffold for bone regeneration. Mol Med Report 17, 5830.Google Scholar
  6. Cao, Z., Wang, D., Li, Y., Xie, W., Wang, X., Tao, L., Wei, Y., Wang, X., and Zhao, L. (2018). Effect of nanoheat stimulation mediated by magnetic nanocomposite hydrogel on the osteogenic differentiation of mesenchymal stem cells. Sci China Life Sci 61, 448–456.PubMedGoogle Scholar
  7. Chen, B., Kankala, R.K., Chen, A., Yang, D., Cheng, X., Jiang, N., Zhu, K., and Wang, S. (2017). Investigation of silk fibroin nanoparticledecorated poly(L-lactic acid) composite scaffolds for osteoblast growth and differentiation. Int J Nanomed 12, 1877–1890.Google Scholar
  8. Chen, C., Liu, Q., Xin, X., Guan, Y., and Yao, S. (2016). Pore formation of poly(ε-caprolactone) scaffolds with melting point reduction in supercritical CO2 foaming. J Supercrit Fluids 117, 279–288.Google Scholar
  9. Choudhury, M., Mohanty, S., and Nayak, S. (2015). Effect of different solvents in solvent casting of porous pla scaffolds—in biomedical and tissue engineering applications. J Biomater Tissue Eng 5, 1–9.Google Scholar
  10. Custódio, C.A., Reis, R.L., and Mano, J.F. (2014). Engineering biomolecular microenvironments for cell instructive biomaterials. Adv Healthc Mater 3, 797–810.PubMedGoogle Scholar
  11. Davies, O.R., Lewis, A.L., Whitaker, M.J., Tai, H., Shakesheff, K.M., and Howdle, S.M. (2008). Applications of supercritical CO2 in the fabrication of polymer systems for drug delivery and tissue engineering. Adv Drug Deliver Rev 60, 373–387.Google Scholar
  12. Delmote, J., Teruel-Biosca, L., Gómez Ribelles, J.L., and Gallego Ferrer, G. (2017). Emulsion based microencapsulation of proteins in poly(Llactic acid) films and membranes for the controlled release of drugs. Polym Degrad Stabil 146, 24–33.Google Scholar
  13. de Matos, M.B.C., Piedade, A.P., Alvarez-Lorenzo, C., Concheiro, A., Braga, M.E.M., and de Sousa, H.C. (2013). Dexamethasone-loaded poly(-caprolactone)/silica nanoparticles composites prepared by supercritical CO2 foaming/mixing and deposition. Int J Pharm 456, 269–281.PubMedGoogle Scholar
  14. de Matos, M.B.C., Puga, A.M., Alvarez-Lorenzo, C., Concheiro, A., Braga, M.E.M., and de Sousa, H.C. (2015). Osteogenic poly(ε-caprolactone)/poloxamine homogeneous blends prepared by supercritical foaming. Int J Pharm 479, 11–22.PubMedGoogle Scholar
  15. Declercq, H.A., Desmet, T., Berneel, E.E.M., Dubruel, P., and Cornelissen, M.J. (2013). Synergistic effect of surface modification and scaffold design of bioplotted 3-D poly-ε-caprolactone scaffolds in osteogenic tissue engineering. Acta Biomater 9, 7699–7708.PubMedGoogle Scholar
  16. Deng, A., Chen, A., Wang, S., Li, Y., Liu, Y., Cheng, X., Zhao, Z., and Lin, D. (2013). Porous nanostructured poly-L-lactide scaffolds prepared by phase inversion using supercritical CO2 as a nonsolvent in the presence of ammonium bicarbonate particles. J Supercrit Fluids 77, 110–116.Google Scholar
  17. Diaz-Gomez, L., Concheiro, A., Alvarez-Lorenzo, C., and García-González, C.A. (2016a). Growth factors delivery from hybrid PCLstarch scaffolds processed using supercritical fluid technology. Carbohyd Polym 142, 282–292.Google Scholar
  18. Diaz-Gomez, L., Yang, F., Jansen, J.A., Concheiro, A., Alvarez-Lorenzo, C., and García-González, C.A. (2016b). Low viscosity-PLGA scaffolds by compressed CO2 foaming for growth factor delivery. RSC Adv 6, 70510–70519.Google Scholar
  19. Duarte, A.R.C., Mano, J.F., and Reis, R.L. (2009). Perspectives on: supercritical fluid technology for 3D tissue engineering scaffold applications. J Bioact Compat Polym 24, 385–400.Google Scholar
  20. Fanovich, M.A., Ivanovic, J., Misic, D., Alvarez, M.V., Jaeger, P., Zizovic, I., and Eggers, R. (2013). Development of polycaprolactone scaffold with antibacterial activity by an integrated supercritical extraction and impregnation process. J Supercrit Fluids 78, 42–53.Google Scholar
  21. Hegde, C., Shetty, V., Wasnik, S., Ahammed, I., and Shetty, V. (2013). Use of bone graft substitute in the treatment for distal radius fractures in elderly. Eur J Orthop Surg Traumatol 23, 651–656.PubMedGoogle Scholar
  22. Hile, D.D., Amirpour, M.L., Akgerman, A., and Pishko, M.V. (2000). Active growth factor delivery from poly(D,L-lactide-co-glycolide) foams prepared in supercritical CO2. J Control Releas 66, 177–185.Google Scholar
  23. Jing, X., Mi, H., Cordie, T., Salick, M., Peng, X., and Turng, L.S. (2014). Fabrication of porous poly(ε-caprolactone) scaffolds containing chitosan nanofibers by combining extrusion foaming, leaching, and freeze-drying methods. Ind Eng Chem Res 53, 17909–17918.Google Scholar
  24. Kankala, R.K., Zhu, K., Li, J., Wang, C., Wang, S., and Chen, A. (2017a). Fabrication of arbitrary 3D components in cardiac surgery: from macro-, micro- to nanoscale. Biofabrication 9, 032002.PubMedGoogle Scholar
  25. Kankala, R.K., Zhang, Y., Wang, S., Lee, C.H., and Chen, A. (2017b). Supercritical fluid technology: an emphasis on drug delivery and related biomedical applications. Adv Healthc Mater 6, 1700433.Google Scholar
  26. Kankala, R.K., Zhu, K., Sun, X., Liu, C., Wang, S., and Chen, A. (2018a). Cardiac tissue engineering on the nanoscale. ACS Biomater Sci Eng 4, 800–818.Google Scholar
  27. Kankala, R.K., Xu, X., Liu, C., Chen, A., and Wang, S. (2018b). 3D-printing of microfibrous porous scaffolds based on hybrid approaches for bone tissue engineering. Polymers 10, 807.PubMedCentralGoogle Scholar
  28. Kankala, R.K., Chen, B., Liu, C., Tang, H., Wang, S., and Chen, A. (2018c). Solution-enhanced dispersion by supercritical fluids: an ecofriendly nanonization approach for processing biomaterials and pharmaceutical compounds. Int J Nanomed 13, 4227–4245.Google Scholar
  29. Kim, H.Y., Kim, H.N., Lee, S.J., Song, J.E., Kwon, S.Y., Chung, J.W., Lee, D., and Khang, G. (2017). Effect of pore sizes of PLGA scaffolds on mechanical properties and cell behaviour for nucleus pulposus regeneration in vivo. J Tissue Eng Regen Med 11, 44–57.PubMedGoogle Scholar
  30. Krause, B., Mettinkhof, R., van der Vegt, N.F.A., and Wessling, M. (2001). Microcellular foaming of amorphous high-T g polymers using carbon dioxide. Macromolecules 34, 874–884.Google Scholar
  31. Lee, S.J., Lee, D., Yoon, T.R., Kim, H.K., Jo, H.H., Park, J.S., Lee, J.H., Kim, W.D., Kwon, I.K., and Park, S.A. (2016). Surface modification of 3D-printed porous scaffolds via mussel-inspired polydopamine and effective immobilization of rhBMP-2 to promote osteogenic differentiation for bone tissue engineering. Acta Biomater 40, 182–191.PubMedGoogle Scholar
  32. Lenas, P., and Ikonomou, L. (2018). Developmental engineering: design of clinically efficacious bioartificial tissues through developmental and systems biology. Sci China Life Sci 61, 978–981.PubMedPubMedCentralGoogle Scholar
  33. Lian, Z., Epstein, S.A., Blenk, C.W., and Shine, A.D. (2006). Carbon dioxide-induced melting point depression of biodegradable semicrystalline polymers. J Supercrit Fluids 39, 107–117.Google Scholar
  34. Luo, G., Huang, Y., and Gu, F. (2017). RhBMP2-loaded calcium phosphate cements combined with allogenic bone marrow mesenchymal stem cells for bone formation. Biomed Pharmacother 92, 536–543.PubMedGoogle Scholar
  35. Mao, J., Duan, S., Song, A., Cai, Q., Deng, X., and Yang, X. (2012). Macroporous and nanofibrous poly(lactide-co-glycolide)(50/50) scaffolds via phase separation combined with particle-leaching. Mater Sci Eng-C 32, 1407–1414.Google Scholar
  36. Mathieu, L.M., Montjovent, M.O., Bourban, P.E., Pioletti, D.P., and Månson, J.A.E. (2005). Bioresorbable composites prepared by supercritical fluid foaming. J Biomed Mater Res 75A, 89–97.Google Scholar
  37. Mathieu, L.M., Mueller, T.L., Bourban, P.E., Pioletti, D.P., Müller, R., and Månson, J.A.E. (2006). Architecture and properties of anisotropic polymer composite scaffolds for bone tissue engineering. Biomaterials 27, 905–916.PubMedGoogle Scholar
  38. Moshiri, A., and Oryan, A. (2012). Role of tissue engineering in tendon reconstructive surgery and regenerative medicine: current concepts, approaches and concerns. Hard Tissue 1, 11.Google Scholar
  39. Nam, Y.S., Yoon, J.J., and Park, T.G. (2015). A novel fabrication method of macroporous biodegradable polymer scaffolds using gas foaming salt as a porogen additive. J Biomed Mater Res 53, 1–7.Google Scholar
  40. Oryan, A., Alidadi, S., Moshiri, A., and Maffulli, N. (2014). Bone regenerative medicine: classic options, novel strategies, and future directions. J Orthop Surg Res 9, 18.PubMedPubMedCentralGoogle Scholar
  41. Park, K.H., Kim, H., Moon, S., and Na, K. (2009). Bone morphogenic protein-2 (BMP-2) loaded nanoparticles mixed with human mesenchymal stem cell in fibrin hydrogel for bone tissue engineering. J Biosci Bioeng 108, 530–537.PubMedGoogle Scholar
  42. Qu, X., Cao, Y., Chen, C., Die, X., and Kang, Q. (2014). A poly(lactide-coglycolide) film loaded with abundant bone morphogenetic protein-2: a substrate-promoting osteoblast attachment, proliferation, and differentiation in bone tissue engineering. J Biomed Mater Res 103, 2786–2796.Google Scholar
  43. Rajabzadeh, S., Liang, C., Ohmukai, Y., Maruyama, T., and Matsuyama, H. (2012). Effect of additives on the morphology and properties of poly (vinylidene fluoride) blend hollow fiber membrane prepared by the thermally induced phase separation method. J Membrane Sci 423–424, 189–194.Google Scholar
  44. Salerno, A., Clerici, U., and Domingo, C. (2014a). Solid-state foaming of biodegradable polyesters by means of supercritical CO2/ethyl lactate mixtures: towards designing advanced materials by means of sustainable processes. Eur Polymer J 51, 1–11.Google Scholar
  45. Salerno, A., Fanovich, M.A., and Pascual, C.D. (2014b). The effect of ethyl-lactate and ethyl-acetate plasticizers on PCL and PCL-HA composites foamed with supercritical CO2. J Supercrit Fluids 95, 394–406.Google Scholar
  46. Salerno, A., Diéguez, S., Diaz-Gomez, L., Gómez-Amoza, J.L., Magariños, B., Concheiro, A., Domingo, C., Alvarez-Lorenzo, C., and García-González, C.A. (2017). Synthetic scaffolds with full pore interconnectivity for bone regeneration prepared by supercritical foaming using advanced biofunctional plasticizers. Biofabrication 9, 035002.PubMedGoogle Scholar
  47. Shen, X., Zhang, Y., Gu, Y., Xu, Y., Liu, Y., Li, B., and Chen, L. (2016). Sequential and sustained release of SDF-1 and BMP-2 from silk fibroinnanohydroxyapatite scaffold for the enhancement of bone regeneration. Biomaterials 106, 205–216.PubMedGoogle Scholar
  48. Singh, L., Kumar, V., and Ratner, B.D. (2004). Generation of porous microcellular 85/15 poly (DL-lactide-co-glycolide) foams for biomedical applications. Biomaterials 25, 2611–2617.PubMedGoogle Scholar
  49. Tomasko, D.L., Li, H., Liu, D., Han, X., Wingert, M.J., Lee, L.J., and Koelling, K.W. (2003). A review of CO2 applications in the processing of polymers. Ind Eng Chem Res 42, 6431–6456.Google Scholar
  50. Tsuji, K., Bandyopadhyay, A., Harfe, B.D., Cox, K., Kakar, S., Gerstenfeld, L., Einhorn, T., Tabin, C.J., and Rosen, V. (2006). BMP2 activity, although dispensable for bone formation, is required for the initiation of fracture healing. Nat Genet 38, 1424–1429.PubMedGoogle Scholar
  51. White, L.J., Hutter, V., Tai, H., Howdle, S.M., and Shakesheff, K.M. (2012). The effect of processing variables on morphological and mechanical properties of supercritical CO2 foamed scaffolds for tissue engineering. Acta Biomater 8, 61–71.PubMedGoogle Scholar
  52. Woodruff, M.A., and Hutmacher, D.W. (2010). The return of a forgotten polymer—polycaprolactone in the 21st century. Prog Polymer Sci 35, 1217–1256.Google Scholar
  53. Xie, Y., Song, W., Zhao, W., Gao, Y., Shang, J., Hao, P., Yang, Z., Duan, H., and Li, X. (2018). Application of the sodium hyaluronate-cntf scaffolds in repairing adult rat spinal cord injury and facilitating neural network formation. Sci China Life Sci 61, 559–568.PubMedGoogle Scholar
  54. Yano, K., Hoshino, M., Ohta, Y., Manaka, T., Naka, Y., Imai, Y., Sebald, W., and Takaoka, K. (2009). Osteoinductive capacity and heat stability of recombinant human bone morphogenetic protein-2 produced by Escherichia coli and dimerized by biochemical processing. J Bone Miner Metab 27, 355–363.PubMedGoogle Scholar
  55. Yang, D., Chen, A., Wang, S., Li, Y., Tang, X., and Wu, Y. (2015). Preparation of poly(L-lactic acid) nanofiber scaffolds with a rough surface by phase inversion using supercritical carbon dioxide. Biomed Mater 10, 035015.PubMedGoogle Scholar
  56. Zeltinger, J., Sherwood, J.K., Graham, D.A., Müeller, R., and Griffith, L.G. (2001). Effect of pore size and void fraction on cellular adhesion, proliferation, and matrix deposition. Tissue Eng 7, 557–572.PubMedGoogle Scholar
  57. Zhao, G., Cao, Y., Zhu, X., Tang, X., Ding, L., Sun, H., Li, J., Li, X., Dai, C., Ru, T., et al. (2017). Transplantation of collagen scaffold with autologous bone marrow mononuclear cells promotes functional endometrium reconstruction via downregulating ΔNp63 expression in Asherman’s syndrome. Sci China Life Sci 60, 404–416.PubMedGoogle Scholar

Copyright information

© Science China Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Hanxiao Tang
    • 1
  • Ranjith Kumar Kankala
    • 1
    • 2
    • 3
  • Shibin Wang
    • 1
    • 2
    • 3
  • Aizheng Chen
    • 1
    • 2
    • 3
    Email author
  1. 1.College of Chemical EngineeringHuaqiao UniversityXiamenChina
  2. 2.Institute of Biomaterials and Tissue EngineeringHuaqiao UniversityXiamenChina
  3. 3.Fujian Provincial Key Laboratory of Biochemical Technology (Huaqiao University)XiamenChina

Personalised recommendations